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Abstract The high-latitude copepod Neocalanus flemingeri exploits the spring phytoplankton bloom to accumulate lipids for survival during food-limited periods and to fuel reproduction. At some point during development, lipid-accumulation ends and pre-adults molt into adults, descend to depth and enter a state of dormancy termed diapause. How and when they determine to make this transition is still unresolved. According to one hypothesis, the trigger is their attaining a threshold amount of lipid fullness. Alternatively, they might follow a genetic program, entering diapause within a narrow developmental window. To better understand the decision, a 5-week laboratory experiment was conducted to assess the effect of food quantity and type on lipid accumulation, biomass and gene expression in N. flemingeri copepodite stage CV. In fed individuals, the initial rate of lipid accumulation slowed by week 5, as a portion of CVs began to molt into adults. While changes in gene expression common to all fed individuals between weeks 1 and 3 were consistent with a developmental program, the duration of the CV stage was variable. Unfed individuals maintained lipid stores initially, suggesting physiological acclimatization to conserve energy. A comparison with gene expression profiles of field-collected individuals suggests similar responses to resources in the environment. 

Abstract BackgroundDiapause is a seasonal dormancy that allows organisms to survive unfavorable conditions and optimizes the timing of reproduction and growth. Emergence from diapause reverses the state of arrested development and metabolic suppression returning the organism to an active state. The physiological mechanisms that regulate the transition from diapause to post-diapause are still unknown. In this study, this transition has been characterized for the sub-arctic calanoid copepodNeocalanus flemingeri, a key crustacean zooplankter that supports the highly productive North Pacific fisheries. Transcriptional profiling of females, determined over a two-week time series starting with diapausing females collected from > 400 m depth, characterized the molecular mechanisms that regulate the post-diapause trajectory. ResultsA complex set of transitions in relative gene expression defined the transcriptomic changes from diapause to post-diapause. Despite low temperatures (5–6 °C), the switch from a “diapause” to a “post-diapause” transcriptional profile occurred within 12 h of the termination stimulus. Transcriptional changes signaling the end of diapause were activated within one-hour post collection and included the up-regulation of genes involved in the 20E cascade pathway, the TCA cycle and RNA metabolism in combination with the down-regulation of genes associated with chromatin silencing. By 12 h, females exhibited a post-diapause phenotype characterized by the up-regulation of genes involved in cell division, cell differentiation and multiple developmental processes. By seven days post collection, the reproductive program was fully activated as indicated by up-regulation of genes involved in oogenesis and energy metabolism, processes that were enriched among the differentially expressed genes. ConclusionsThe analysis revealed a finely structured, precisely orchestrated sequence of transcriptional changes that led to rapid changes in the activation of biological processes paving the way to the successful completion of the reproductive program. Our findings lead to new hypotheses related to potentially universal mechanisms that terminate diapause before an organism can resume its developmental program. 

Abstract Many arthropods undergo a seasonal dormancy termed “diapause” to optimize timing of reproduction in highly seasonal environments. In the North Atlantic, the copepodCalanus finmarchicuscompletes one to three generations annually with some individuals maturing into adults, while others interrupt their development to enter diapause. It is unknown which, why and when individuals enter the diapause program. Transcriptomic data from copepods on known programs were analyzed using dimensionality reduction of gene expression and functional analyses to identify program-specific genes and biological processes. These analyses elucidated physiological differences and established protocols that distinguish between programs. Differences in gene expression were associated with maturation of individuals on the reproductive program, while those on the diapause program showed little change over time. Only two of six filters effectively separated copepods by developmental program. The first one included all genes annotated to RNA metabolism and this was confirmed using differential gene expression analysis. The second filter identified 54 differentially expressed genes that were consistently up-regulated in individuals on the diapause program in comparison with those on the reproductive program. Annotated to oogenesis, RNA metabolism and fatty acid biosynthesis, these genes are both indicators for diapause preparation and good candidates for functional studies.